Molecular Mechanism of MicroRNA-200c Regulating Transforming Growth Factor-β (TGF-β)/SMAD Family Member 3 (SMAD3) Pathway by Targeting Zinc Finger E-Box Binding Homeobox 1 (ZEB1) in Hypospadias in Rats

Abstract

BackgroundThe aim of this study was to explore effects of microRNA-200c regulating TGF-β/Smad3 pathway by targeting Zeb1 on the occurrence and development of hypospadias and to evaluate the relationship between microRNA-200c and occurrence of hypospadias.Material/MethodsPregnant rats with a gestational age of 12 days were allocated into 2 groups; one received gavage of DEHP-contained soybean oil (1 ml/day, 8 days; Group A) and the other had gavage of normal soybean oil (1 ml/day, 8 days; Group B). Baby rats with hypospadias from Group A were assigned to the model group (n=20) and healthy baby rats from Group B were assigned to the control group (n=20). Real-time quantitative polymerase chain reaction (qRT-PCR), immunohistochemistry and Western blot analysis were performed to detect microRNA-200c, Zeb1, TGF-β, and Smad3 mRNA and protein expressions in the model group (n=20) and the control group (n=20). The relationship between microRNA-200c and Zeb1 was detected using a dual-luciferase reporter gene experiment. After the in vitro intervention experiment in fetal rat penises, Western blot was used to detect the expression of Zeb1, TGF-β, and Smad3.ResultsIn the model group, microRNA-200c was expressed at a low level, and microRNA-200c expression in control group was 2.1 times higher than in the model group (P<0.05). When compared with the control group, mRNA expressions, protein expressions, and positive rates of Zeb1, TGF-β, and Smad3 were higher in the model group (all P<0.01). Luciferase gene report determined that Zeb1 is a target gene of microRNA-200c. The in vitro intervention experiment in fetal rat penises found that a high concentration of microRNA-200c inhibited hypospadias occurrence by suppressing the expression of Zeb1, TGF-β, and Smad3.ConclusionsMicroRNA-200c was expressed in hypospadias penis tissues at low levels and was negatively correlated with Zeb1 expression. MicroRNA-200c up-regulated Zeb1 expression to regulate the TGF-β/Smad3 pathway, which led to the occurrence of hypospadias.

Potential risk factors for hypospadias and negative correlation with DICER1 (rs3742330) A>G variant in Algerian population: A case‐control study

“…One study revealed a notable reduction in microRNA‐200c expression in hypospadias penile tissues (Qian et al, 2016). Additionally, another study suggested miR‐494's potential role in hypospadias pathogenesis (Tian et al, 2020).…”

“…In recent studies, there has been an exploration of the functions and roles of microRNAs (miRNAs) in the developmental processes of reproductive systems. However, only a limited number of investigations have specifically concentrated on the incidence and progression of hypospadias in male infants (Deng et al, 2021; Qian et al, 2016; Tian et al, 2020).…”

Perfil de expressão plasmática dos microRNAs miR-200c e miR-210 em pacientes portadores de diferenças do desenvolvimento sexual (DDS) 46,XY de causa indeterminada

“…Estudos de miRNAs em órgãos do sistema genital masculino A presença de hipospádia é um achado frequente em pacientes com DDS46,XY e resulta da ação androgênica deficiente com consequente subvirilização genital. Para avaliar a influência dos miRNAs nesta condição de anormalidade da virilização genital,Qian (2016) realizou uma análise da literatura e selecionaram o miR-200c como candidato, baseados nos alvos validados para este miRNA. SMAD3, fatores importantes que, em conjunto, tem um papel no desenvolvimento da fibrose uretral.…”